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Please use this identifier to cite or link to this item: http://repository.iitr.ac.in/handle/123456789/1079
Title: A synthetic luxCDABE gene cluster optimized for expression in high-GC bacteria
Authors: Craney A.
Hohenauer T.
Xu Y.
Navani, Naveen Kumar
Li Y.
Nodwell J.
Published in: Nucleic Acids Research
Abstract: The luxCDABE operon of the bioluminescent bacterium Photorhabdus luminescens has proven to be a superb transcriptional reporter. It encodes a luciferase (LuxA and LuxB) and the enzymes that produce its substrate (LuxC, LuxD and LuxE) so cells that express the cluster emit the 490-nm light spontaneously. The sequence of genes is AT-rich (>69%) and for this and other reasons, they are not expressed efficiently in high-GC bacteria like Streptomyces coelicolor. We therefore constructed a synthetic luxCDABE operon encoding the P. luminescens Lux proteins optimized for expression in high-GC bacteria. We tested the genes using transcriptional fusions to S. coelicolor promoters having well-established expression profiles during this organism's life cycle. The hrdB gene encodes a housekeeping sigma factor; while ramC is important for the formation of the spore-forming cells called aerial hyphae and whiE is required for the production of a grey, spore-associated pigment that is deposited in the walls of developing spores. Using these fusions we demonstrated that our synthetic lux genes are functional in S. coelicolor and that they accurately report complex developmental gene expression patterns. We suggest that this lux operon and our procedure for generating synthetic high-GC genes will be widely useful for research on high-GC bacteria. © 2007 The Author(s).
Citation: Nucleic Acids Research (2007), 35(6): -
URI: https://doi.org/10.1093/nar/gkm086
http://repository.iitr.ac.in/handle/123456789/1079
Issue Date: 2007
ISSN: 3051048
Author Scopus IDs: 16300695400
16301365400
7406450578
6507073924
8082236600
6701756918
Author Affiliations: Craney, A., Department of Biochemistry and Biomedical Sciences, McMaster University, Health Sciences Centre, 1200 Main Street W Hamilton, Hamilton, ON L8N 3Z5, Canada
Hohenauer, T., Department of Biochemistry and Biomedical Sciences, McMaster University, Health Sciences Centre, 1200 Main Street W Hamilton, Hamilton, ON L8N 3Z5, Canada
Xu, Y., Department of Biochemistry and Biomedical Sciences, McMaster University, Health Sciences Centre, 1200 Main Street W Hamilton, Hamilton, ON L8N 3Z5, Canada
Navani, N.K., Department of Biochemistry and Biomedical Sciences, McMaster University, Health Sciences Centre, 1200 Main Street W Hamilton, Hamilton, ON L8N 3Z5, Canada
Li, Y., Department of Biochemistry and Biomedical Sciences, McMaster University, Health Sciences Centre, 1200 Main Street W Hamilton, Hamilton, ON L8N 3Z5, Canada
Nodwell, J., Department of Biochemistry and Biomedical Sciences, McMaster University, Health Sciences Centre, 1200 Main Street W Hamilton, Hamilton, ON L8N 3Z5, Canada
Funding Details: We are grateful to Dr John Capone who supported this project as Associate Dean of Medicine at McMaster University. We also thank the staff of the MOBIX centre for their patient assistance with the construction of oligonucleotides and the sequencing of the synthetic lux genes and Marie Elliot for providing the ramR null strain and its parental strain M600. This work was supported with grants MOP-57684 and MOP-68817 and a New Investigator Award for JN from the Canadian Institutes for Health Research. Funding to pay the Open Access publication charge was provided by CIHR.
Corresponding Author: Nodwell, J.; Department of Biochemistry and Biomedical Sciences, McMaster University, Health Sciences Centre, 1200 Main Street W Hamilton, Hamilton, ON L8N 3Z5, Canada; email: nodwellj@mcmaster.ca
Appears in Collections:Journal Publications [BT]

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